不同着色期荔枝果皮总RNA的提取及mRNA差显分析

以妃子笑和糯米糍2个主栽荔枝品种不同着色期的果皮为试材,经过大量的对比和筛选,采用改良Bugos法提取果皮中的总RNA,经核酸蛋白测定仪测定和凝胶电泳显示提取的总RNA完整性强,质量好,产率高(最高可达318.7μg/g),可用于RT-PCR等分子生物学实验。进一步的DDRT-PCR分析表明:高分辨力的cDNA片段在500～2 000 bp之间,不同品种、不同着色期的荔枝果皮,基因表达有较大差异。

Total RNA extraction and mRNA differential display analysis of litchi pericarps at different coloring stages

The pericarp of litchi shows close relationship with fruit cracking resistance,pigmentation,post-harvest browning and so on,and therefore has been the highlight of fruit studies.With the researches advanced,it is essential to illuminate the physiological mechanism of pericarpe by means of molecular studies.The high quality RNA,however,was very difficult to extract from the tissue of litchi pericarp due to easily browning and low content,and subsequently this may retard molecular physiological researches in the pericarp of litchi.In the present study,total RNA of litchi pericarp of 2 cultivars,Feizixiao and Nuomici,at different coloring stages was extracted with improved Bugos method.The total RNA was proved to be relative intact and good quality,and the content of total RNA was high(the highest one reached 318.7 μg/g) tested by nucleic acid/protein assay and gel electrophoresis.And hence the extracted RNA could be used in molecular biology experimentation,such as RT-PCR etc.The results of DDRT-PCR analysis showed that the cDNA bands of highly-resolving power existed among 500 bp and 2 000 bp,and there displayed marked difference in the gene expression in the pericarps of different litchi cultivars at different coloring stages.